Abstract: Abstract: Expression vectors are important materials in establishment of plant transformation system. In this study, we constructed a GFP expression vector pGWB14-GFP through Gateway cloning technology. Then we transiently transformed this vector into Onion peels and wheat leaves by gene gun method and GFP expression was detected by epi-fluorescence microscope. The results showed that GFP can function as usual in cells of both onion and wheat, suggesting this vector can be used in the transformations of both dicot and monocot plants. This vector cotains GFP expression cassette and selection markers for kanamycin, and can be used in both dicot and monocot plants. With these advantages, this vector would become a powerful tool for the establishment of plant transformation system.